Preservation of food products



- 'Patenteci Apr. 24, 1951 1 UNITED; )STATE PRESERVATION pr noonPRODUCTS Lloyd 5B. Jensen, Chicago; and William A. Miller, EvergreenPark, Ill., as'signors to Swift & Com? pany, Chicago, 111., acorporation of Illinois i No Drawing. Applicationflctoherfi,1946,

Serial No. 701,406 Claims. (-01. 291-51 6 The present invention relatesto the preparaime or an antibio ic ubstan e and it has to do mere tiu1ar1y. withthe sen e ttai n f an antimicrobial ,substanceofplantorigin.

I An object of t e p esent i vent on s to p 7 videa potent antibiotic s1,1 -bstance of plant ,origin.

A fu13ther object of the invention is to provide an tibi e u et ncenw hi a -t n? i llul anr piv .,An additional object ofethe invention is toprovide an antibiotic substance which isheat stable.

Yet another object of ,the invention is to .employan antibioticsubstance of grapevine origin as an effective p ese va ve fo $0001 ,prdu Qther objects and. advanta es .of the invention will be apparent.from ,a descrip ion of the in vention given below. l

The present invention-contemplatesthe preparati of an antibi t c sustanc n tu a ais m ciated with tissues of the members of the ,plantgenus Vitis, commonlyknown as grape, fWehafve e l-1 d th t p ci ll th tvated g ap v ne, including, the root, yields an: antibiotic substanceofggreat'jpotency as ,hereinaftermore .fullyqde scribed- 1 W hav also lfun tha the a i ti substance produced from the grapevine is lethal tothose microorganisms 'laifgelyjespqnsible for clostridial and eilla i ffold slidilaeeand h ococeus ood poi onin f l eiantib t c m teri l prepaed from the .grapevin 'may, therefo e b advan a eou l u ed't prem e odpro u j The ant b ot c sub t n of t e p esen 111N311? tion 'hasbeenfoundto destroy as '-forniing;bacilli of, the genus ,Badillusis a' 'fial u el (var. m'ger) ;the bacilli ofthegenusClostridium,

such as Clostridiym spovjogene c, andrthe diseaseproducingbacilli of thegenus 'Bru'cella, such as Brucella suis; the staphylococci,suchfasiSftaphzi- *Zoco'cculs au'reu's. Weha ve 'also found our antibiotic substance to belethalj against the bacterial.

ascertain theirsponse 0f all the existing species anetmeues'ormicroorganisms to the herein described antibiotic material, but theeffects ofthis material-"againstthe microorganisms 'l ereinabovementioned illustrate the comniereiakand medieal valuesthere'of: 1 i

#10 species Achromobacterpemlem,:the orga-nismre sqlun the in :5

"extract were wmapleitl the "hint,

c'ntain eelt thereunde v illu obtaine in aoco examples- 0 c 1 :1 1 all IExample lei-5 0 ams 19f eranet ne ammamately two to three years old, wasfinely e51.

ground woodwas plane in a last and 299 red. with .lacet ne. eflas was1116, 5! ,rQQm tem eratures for in means and shaken a er vals. .iDhelacetonezlayer. containing the acetone soluble" constituents lois'the.grapevine wassena: rated efromudthe insoluble, materialmy filtration,and the -acetone removed iromitheextractibyldise tillation sunder 1reduced .pressure :at A0 L; -il he acetone-free ex-tract; ahighlafiviscous :mass, was

taken up-in- '50 ml'; of ethyl alcohol, -so that 1 ml.

of -the alcoholic extract contained :the acetone soluble material in 3-gralns o f grapevine (l-mll=3 'g.' oi-vine). h

In order to evaluate the effectiveness of the antibiotic material in thealcohol solution obtained in accordance witliilExample I, the followingtest was performed: 1

Serial dilutions of the e'xtract (l ml.=3 g. of wood) were made .in 10,9broth tube and the tuha nwunta ti h taqts ia in lu qem i 10 110 l to450,000 and inocglatedm th la ops of -Bel-1 10111";broth cultures of afood noisgninlg strain gOf fitaphylocpcozgs aureu'e (50,000 cells),Bacillus niger, clostrzldium'sz'iorogenes and Achro- .mohee ememleme llestubeseuere tlienlallew toincubatecat a itempet ture -Q-f :i'3i UC-@1139: in-

TABLE I Grapeuiue'extract (1 ml.=3 gms. of pine) Dilutions in brothOrganism Days 100 500 1,000 2,000 5,000 10,000 20,000 40,000 1 Staph.aureus 2 a v r 6 ;'1- Bacillus mger 2 6 I 1 ('lostndm'm sporooeues I i 1Achromobacter perolens 2 6 l Grown in Difco peptone colloid medium.

The data given in the above table indicate that Although acetone is thepreferred primary solthe antibiotic efiects of the grapevine extractvent, we have found that other organic solvents vary in potency whentested against different are suitable, for example, dioxane, ethylacetate, organisms. Thus, the extract kills Staphylococmethyl ethylketone and ethanol. We have also cus in dilutions of 1 to.1,000,,Bacillus niger in 25 found our antibiotic material to be slightlysoludilutions of 1 to 500, and 1 to 100 for Clostridium ble in water. Ysporogeues and Achromobacter peroleus. The It is obvious that manyvariations in the solbactericidal properties of the extract, lethallyvent treating process may be advantageously effective in the dilutionsindicated for the various employed. For example, since the antibioticorganisms, were demonstrated by inoculating material isolated fromgrapevine is heat-stable, nutrient agar plates with 0.1 cc. of theaforesaid solvent temperatures may range from room temdilutions whichhad been allowed to incubate for peratures to boiling temperature of thesolvent, six days. After three days incubation of the with consequentvariations in extraction time, inoculated plates no viable bacteria wereobthat is, the higher the temperature, generally the served as a resultof the transfers. The test inless time required for extraction.Moreover, the dicates that the bacteria had been killed indiluextraction process may be carried out in one step, tions of 1 to1,000 for Staphylococcus aureus, 1 to for example, with ethanol or witha mixture of 500 for Bacillus m'yer, and 1 to 100 for both solvents. Wehave found it preferable to process Clostridium sporogenes andAchromobacter perthe antibiotic substance contained in the grapeolens.40 vine by a two-stage solvent process involving Example II .-50 gramsof cultivated grapevine, first, treating the substance with a primarysolabout 10 years old, which was beginning to sap, vent which issubsequently removed and then, was finely ground, covered with acetoneina flask second, taking up the resulting extract with anand held at 20 C.for three days with occasional other solvent, so as to form a solution,which is a shaking. The acetone was taken 01f at 41 C. by 45 convenientform of using the product. We have means of a water pump, and theresidue dissolved found that acetone is a very effective primary in ml.of ethanol so that 1 ml. of alcoholic solvent in producing theantibiotic. substance extract contained the acetone-soluble material infrom the plant tissue. After evaporating ofi the one g. of wood (1 ml.=1g. of wood). acetone the preferred solution is made with eth- Dilutionsof the extract were made in nutrient 50 anol. By using alcohol weutilize its well-known broth and each series of dilutions wereinocuproperties, such as non-toxicity and ability to lated with 24-hourcultures of Staphylococcus prevent contamination. aureus, Bacillus nigerand Brucella suis, with the In the processing of food products a seriousresults tabulated in Table II. problem is that of food preservation, andal- 1 though great strides have been made in improv- TABLE II ing thequality and keeping time of various food products by refrigeration,canning, curing, plant Dilutions in broth sanitation, etc, the problemof the preservation of food requiresthe constant attention of the oodprocessor to improve long-established methorgansm ow'wl ods of treatingfood and to devise new means of 1 mus I food control.

Ba l u y v. Food spoilage is due principally to microbial action, andthe species of microorganisms responsible for food deterioration arenumerous. The

The tubes representing serial dilutions from a sporing generaClostridium and Bacillus, for minimum dilutio of 1 to 100 to a maximum(11.7 ample, when contaminating food, cause food lution of 1 to 2,000were allowed to incubate at spoilage- Another Serious concern of thefood 37 c; At-th nd of five days th was no handler is food poisoning,which is often of an dence of bacterial growth, as indicated by theminus sign,'for Staphylococcus aureus in dilutions of 1 to 1,000, 1 to500 for Bacillus uiger, and 1 to 400 for Brucella suis. .The positivesigns indicate living bacteria.

insidious nature because food products contaminated with harmfulbacteria often appear organoleptically sound. Among thefood poisoningbacteria,,the most common food poisoner is the staphylococcus. Inaddition to foodspoilage and by the Brucella organisms:

food. use of; disease,- for example, briiceilosi-s produced Theantibacterial substance of the resent iriveiitiori may be used to retardor prevent microbial growth in various food products. This material mayadvantageously be employed in treating meat products such as "hair-isausages and canned iiieats'; filled pastrieisiich as cream Duffs,chocolate eclairs, custards, etc.; and canned vege-r ta es, to, preventsweuings cue, for example; to organisms or the genus Baeiuus. 1 I 1 Theinvention will be more fullyfuiiderstood from the following descriptionof a method of using the antibiotic substance, in connection with thecuring of hams.

' Within recent years there has been developed a curing process formeats and particularly hams 7 known as a quick-cure which requires onlya fraction of the time whichwas normally required by the old processes.Essentially the quick-cure process consists in pumping the picklingsolution under pressure into the vascular system of the meat. .By thismeans a ham can be cured in a period of time ranging from to 18 days,rather than the 60 to 90 days required by the old processes in which themeats were usually immersed in a bath of the pickling fluid.

There has arisen, however, as the result of extensive commercial useofthe quick-curing method an attendant serious health problem. The

hams and other meats cured by the older immersion processes wereextremely resistant to the development of pathogenic bacteria, of whichvarious representatives of the Staphylococci may be mentioned astypical, and the general public has come to rely on this property in itseating and storage habits. Hams processed by quick-curing, on the otherhand, have resulted in a meat product which, though highly satisfactorywhen fresh, is often less resistant to these organisms, and cases offood poisoning have resulted frOm improper handling due to unfamiliaritywith this fact. 7

As hereinbefore mentioned, we have found that the antibiotic reagentherein described is lethal to a number of microbes in a 1 to 1,000dilution of the extract, the extract being of such strength that 1 ml.of the alcoholic solution thereof con- 7 tained the acetone-solubleconstituents in 3 grams of wood. Therefore, the addition of 1 part ofthe solution to 1,000 parts of the conventional pickling solution willresult in a meat product cured therewith which has a very markedincrease in resistance to the development of pathogenic organisms.

7 lbs. sodium nitrate .10 oz. sodium nitrite lbs. sugar 378.5 cc.antibiotic solution The following experiments will further mustrate thepractical application of my invention.

An amount of pickle was inoculated with a large number ofa foodoiso'ningtype of Staphylococcus aureus organisms which would representthe most unfavorable conditions which could possibly be encountered. Thepickle was then separated into two equal portions, and to one portionsufficient amount of grapevine product was added toform' alto 1000dilution of the product. To the other portion of pickle no antibioticsubstance was added. Twelve hams were then'ciired by the quickcuring'methodsix of themwith pickle containing the antibiotic substance and sixwith pickle without the substance. The .hams were then smoked accordingto ordinary processes. The hams were incubated at 99 F. and at the endof two weeks the six hams containing the antibiotic substance did notharbor viable staphylococci, while the other hams showed the, presenceof viable staphylococci.

Luncheon meat was cured according to conventional methods in which theantibiotic substance was present in a lto 100,0 dilution. The meatproduct was placed in six 6-pound cans and retorted to an insidetemperature of F. and incubated at 99 F. After 30 days of incubation, noswelling of the cans could be observed. The control cans swelled in oneWeek.

Obviously, many modifications and variations of the invention, ashereinabove set forth, may be made without departing from the spirit andscope thereof, and therefore only such limita tions should be imposed asare indicated in the appended claims.

We claim:

1. In the process of treating food products, the step of adding to afood product a small amount of an alcoholic solution of antibioticsubstance derived from the wood of the members of the genus Vitis topreserve said food products against pathogenic bacteria responsible forfood poisoning and spoilage.

2. A meat pickle including a small amount of an antibiotic substance ofthe wood of the members of the genus Vitis extracted from the said woodby a non-aqueous organic solvent selected from the group consisting ofacetone, dioxane, ethylacetate, methylethylketone, and ethanol.

3. In the process of quick-curing meats wherein a pickling solution isinjected into said meat, the step of adding to said pickling solution asmall amount of the antibiotic substance derived from the wood of themembers of the genus Vitis by treatment with a non-aqueous organicsolvent, whereby the growth of pathogenic bacteria responsible for foodspoilage and poisoning in the meat are substantially retarded after thesaid solution is injected into the meat.

4. In the process of treating food products, the step of adding to afood product a small amount of an antibiotic substance derived from thewood of the members of the genus Vitis by treatment with a non-aqueousorganic solvent to preserve said food products against pathogenicbacteria responsible. for food poisoning and spoilage.

5. A meat pickle including a small amount of an antibiotic substance ofthe wood of the members derived from the genus Vitis by treatment with anon-aqueous organic solvent.

LLOYD B. JENSEN. WILLIAM A. MILLER.

(References on following page) REFERENCES CITED UNITED STATES PATENTSName Date Whiteley Apr. 30, 1872' Chesebrough Mar. 1, 1892 Taub et a1.Oct. 13, 1914 Krebs -g. Sept; 1, 1925 Masucci Oct. 27, 1925 Paddock June'22, 1937 Schertz et a1; Nov. 2, 1937 Number 2,117,478 2,173,9922,180,750

OTHER REFERENCES 7 Natu're, May 13, 1944, vol. 153, page 598, articleentitled Antibacterial Substances in Green 10 Plants.

Chemical and Engineering'News, September 1945, page 1622', articleentitled Penicillin as a Preseryative.

1. IN THE PROCESS OF TREATING FOOD PRODUCTS, THE STEP OF ADDING TO AFOOD PRODUCT A SMALL AMOUNT OF AN ALCOHOLIC SOLUTION OF ANTIBIOTICSUBSTANCE DERIVED FROM THE WOOD OF THE MEMBERS OF THE GENUS VITIS TOPRESERVE SAID FOOD PRODUCTS AGAINST PATHOGENIC BACTERIA RESPONSIBLE FORFOOD POISONING AND SPOILAGE.